同种异体PRP联合ADSC对小鼠皮肤创面再生愈合情况及血清HA、VEGF水平的影响

Effects of allogeneic PRP combined with ADSC on wound healing and serum HA and VEGF levels in mice

目的:探究同种异体富血小板血浆(PRP)联合脂肪源性间充质干细胞(ADSC)对小鼠皮肤创面再生愈合情况及血清透明质酸(HA)、血管内皮生长因子(VEGF)水平的影响。方法:45只小鼠随机分为3组(n=15),分别为对照组(control)、ADSC组和ADSC+PRP组。3组小鼠均建立创面损伤模型,control组皮下注射生理盐水,ADSC组皮下注射同种异体PRP,ADSC+PRP皮下注射ADSC和PRP。分别在建模后3、5、7d观察创面愈合情况以及用HE染色观察组织形态学特点。使用Westernblot检测CollagenI和CollagenⅢ水平。检测并比较各组血清HA、VEGF水平。结果:建模后3、5、7d时ADSC组和ADSC+PRP组的创面愈合率显著高于control组(P<0.01),ADSC+PRP组的创面愈合率显著高于ADSC组(P<0.01)。建模后3d和5d时control组和ADSC组的炎性反应明显,ADSC组有少量肉芽组织,而ADSC+PRP组的炎症反应较轻,并且有较多的肉芽组织;建模后7dADSC+PRP组新生血管较多并且再上皮化率较高,而control组和ADSC组的上皮化较差,并且新生血管较少。建模后3、5、7d时ADSC组和ADSC+PRP组的CollagenI和CollagenⅢ表达水平显著高于control组(P<0.01),ADSC+PRP组的CollagenI和CollagenⅢ显著高于ADSC组(P<0.01)。建模后3、5、7d时ADSC组和ADSC+PRP组的血清HA和VEGF显著高于control组(P<0.01),ADSC+PRP组的血清HA和VEGF显著高于ADSC组(P<0.01)。结论:同种异体PRP联合ADSC可促进小鼠皮肤创面再生愈合,其机制可能与上调VEGF和HA的表达,促进血管和肉芽组织的生成有关。

Objective:elet-rich plasma(PRP)combined with adipose-derived mesenchymal stem cells(ADSC)on skin wound healing and serum hyaluronic acid(HA)and vascular endothelial growth factor(VEGF)levels in mice.Methods:Forty-five mice were randomly divided into three groups(n=15),which were control,ADSC,and ADSC+PRP.The wound injury model was established in all three groups.The control group was injected subcutaneously with normal saline,the ADSC group was injected subcutaneously with allogeneic PRP,and the ADSC+PRP was injected subcutaneously with ADSC and PRP.Wound healing was observed at 3d,5d,and 7d after modeling,and histomorphological features were observed by HE staining.Collagen I and CollagenⅢlevels were detected using Western blot.The levels of serum HA and VEGF in each group were detected and compared.Results:The wound healing rate of ADSC group and ADSC+PRP group was significantly higher than that of control group at 3d,5d and 7d after modeling(P<0.01).The wound healing rate of the ADSC+PRP group was significantly higher than that of the ADSC group(P<0.01).The inflammatory response of the control group and the ADSC group was significant at 3 and 5 days after modeling,and there was a small amount of granulation tissue in the ADSC group.The ADSC+PRP group had a milder inflammatory response and more granulation tissue;After 7d,the ADSC+PRP group had more neovascularization and higher re-epithelialization rate.The control group and the ADSC group had poorer epithelialization and fewer new blood vessels.The expression levels of Collagen I and CollagenⅢin ADSC group and ADSC+PRP group were significantly higher than those in control group at 3d,5d and 7d after modeling(P<0.01).Collagen I and CollagenⅢin the ADSC+PRP group were significantly higher than those in the ADSC group(P<0.01).Serum HA and VEGF in ADSC group and ADSC+PRP group were significantly higher than those in control group at 3d,5d and 7d after modeling(P<0.01).Serum HA and VEGF were significantly higher in the ADSC+PRP group than in the ADSC group(P<0.01).Conclusion:Allogeneic PRP combined with ADSC can promote the regeneration and healing of skin wounds in mice.The mechanism may be related to the up-regulation of the expression of vascular endothelial growth factor and HA and the promotion of the formation of blood vessels and granulation tissue.